Manchester United trail the Spanish giants in second position having moved up the log, with Bayern Munich and Barcelona filling in at third and fourth positions respectively.
According to a report released by accountancy firm Deloitte, Manchester United's income of £433million in 2013-14 left them behind Real Madrid, who brought in £459m, but ahead of third-placed Bayern Munich (£407.7m), Barcelona in fourth (£405.3m) and Paris Saint-Germain in fifth (£396.6m). Real Madrid have now been the richest club by earnings for 10 years in a row.
Manchester City (£346.5m), Chelsea (£324.4m), Arsenal (£300m) and Liverpool (£255.8m) are next in the top 10. Newcastle and Everton squeeze into the top 20, with West Ham and Aston Villa just below them.
Southampton are the 25th richest club in the world (on £106.1m), with Sunderland, Swansea and Stoke inside the top 30.
The rest of last season's Premier League teams earned more than the £80m necessary for a top-40 place. This was largely down to lucrative new 2013-16 TV deals which began to pour into club coffers last season.
Austin Houlihan, senior manager at Deloitte, said: "This is testament to the huge appeal of the Premier League globally and the equality of distribution the clubs enjoy relative to their European counterparts."
Top 30 Richest football clubs by revenue (2013-14)
1. Real Madrid - £459.5m
2. Manchester United - £433.2m
3. Bayern Munich - £407.7m
4. Barcelona - £405.2m
5. Paris Saint-Germain - £396.5m
6. Manchester City - £346.5m
7. Chelsea - £324.4m
8. Arsenal - £300.5m
9. Liverpool - £255.8m
10. Juventus - £233.6m
11. Borussia Dortmund - £218.7m
12. AC Milan - £208.8m
13. Tottenham - £180.5m
14. Schalke 04 - £178.9m
15. Atletico Madrid - £142.1m
16. Napoli £137.8m
17. Inter Milan - £137.1m
18. Galatasaray - £135.4m
19. Newcastle United - £129.7m
20. Everton £120.5m
21. West Ham United - £105.3m
22. Aston Villa - £101.9m
23. Marseille - £100m
24. Roma - £97.7m
25. Southampton - £97.3
26. Benfica - £96.6
27. Sunderland - £95.7m
28. Hamburg - £92.2m
29. Swansea City - £90.5m
30. Stoke City - £90.1m